# US 10,308,965 B2 · Biological oils and production and uses thereof

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- Number: US 10,308,965 B2
- Document ID: US10308965B2
- Status: Granted
- Kind: B2
- Application number: 15/237,988
- Filed: 2016-08-16
- Published: 2016-12-08
- Granted: 2019-06-04
- Earliest claimed: 2007-09-12
- Assignees: Dsm IP Assets B.V.
- Applicants: DSM IP Assets B.V.
- Inventors: James Casey Lippmeier; Joseph W. Pfeifer, III; Jon Milton Hansen, III; Kirk E. Apt; William Robert Barclay; Paul Warren Behrens; David Christian Martin
- Web page: https://patentgrep.com/patent/US10308965B2

## CPC

- C12P 7/6458: by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
- C10L 1/02: essentially based on components consisting of carbon, hydrogen, and oxygen only
- C10L 1/026: {for compression ignition}
- C12P 7/6472: containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
- C12P 7/649: Biodiesel, i.e. fatty acid alkyl esters
- C10L 2200/0469: Renewables or materials of biological origin
- C10L 2270/026: for diesel engines, e.g. automobiles, stationary, marine
- C10L 2270/04: for turbines, planes, power generation
- C10L 2290/26: Composting, fermenting or anaerobic digestion fuel components or materials from which fuels are prepared
- Y02E 50/10: Biofuels, e.g. bio-diesel
- Y02P 30/20: using bio-feedstock
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C10L: FUELS NOT OTHERWISE PROVIDED FOR; NATURAL GAS; SYNTHETIC NATURAL GAS OBTAINED BY PROCESSES NOT COVERED BY SUBCLASSES C10G OR C10K; LIQUIFIED PETROLEUM GAS; USE OF ADDITIVES TO FUELS OR FIRES; FIRE-LIGHTERS
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The present invention provides biological oils and methods and uses thereof. The biological oils are preferably produced by heterotrophic fermentation of one or more microorganisms using cellulose-containing feedstock as a main source of carbon. The present invention also provides methods of producing lipid-based biofuels and food, nutritional, and pharmaceutical products using the biological oils.

## Claims

### Claim 1

A method for producing a biological oil comprising
(a) growing an oil-producing microorganism of the kingdom Stramenopile by heterotrophic fermentation in a medium comprising cellulose as a carbon source, and
(b) isolating or purifying the biological oil made in step (a); wherein about 11% to about 99% of unsaturated fatty acids in said biological oil are polyunsaturated fatty acids, and wherein said fermentation is performed in a fermentor selected from the group consisting of fiber reinforced polymer fermentors, metal matrix composite fermentors, ceramic matrix composite fermentors, thermoplastic composite fermentors, metal fermentors, epoxy lined carbon steel fermentors, plastic lined carbon steel fermentors, plastic fermentors, fiberglass fermenters, and concrete fermentors.

### Claim 2 (depends on 1)

The method of claim 1, wherein greater than about 50% of unsaturated fatty acids in said biological oil are polyunsaturated fatty acids.

### Claim 3 (depends on 1)

The method of claim 1, wherein said microorganism is a Thraustochytrid.

### Claim 4 (depends on 1)

The method of claim 1, wherein said microorganism is selected from the group consisting of microorganisms of the genus Schizochytrium, microorganism of the genus Thraustochytrium, and microorganisms of the genus Ulkenia.

### Claim 5 (depends on 1)

The method of claim 1, wherein said microorganism saccharifies said cellulose.

### Claim 6 (depends on 1)

The method of claim 1, wherein said microorganism is resistant to or degrades feedstock components selected from the group consisting of lignin, hemicellulose, plant oil, plant extracellular polysaccharides, and combinations thereof.

### Claim 7 (depends on 1)

The method of claim 1, wherein said microorganism is a genetically modified microorganism.

### Claim 8 (depends on 1)

The method of claim 1, wherein said microorganism produces oil in triglyceride form in an amount of from about 25% to about 85% by weight of its dry biomass.

### Claim 9 (depends on 1)

The method of claim 1, further comprising performing autolysis or induced lysis of said microorganism after said microorganism has produced oil in an amount of from about 30% to about 90% by weight of its dry biomass.

### Claim 10 (depends on 1)

The method of claim 1, further comprising inducing lysis of said microorganism by exposing said microorganism to a condition favorable for lysis selected from the group consisting of a pH, a temperature, the presence of an enzyme, the presence of a detergent, physical disruption, and combinations thereof.

### Claim 11 (depends on 1)

The method of claim 1, wherein said medium comprising cellulose comprises a source of cellulose selected from the group consisting of grass, sugar cane, agricultural waste, waste paper, sewage, wood, an organism of the kingdom viridiplantae, and combinations thereof.

### Claim 12 (depends on 1)

The method of claim 1, wherein said fermentation is performed in a nonsterile fermentor.

### Claim 13 (depends on 1)

The method of claim 1, wherein said fermentation is carried out in a fermentor that is submerged in water.

### Claim 14 (depends on 1)

The method of claim 1, wherein said fermentation is carried out in fermentors having cooling systems connected in series such that cooling water effluent from a first fermentor or a set of fermentors in the series is used as a cooling water supply for a second fermentor or set of fermentors in the series.

### Claim 15 (depends on 1)

The method of claim 1, wherein said fermentation is carried out in fermentors having gas systems connected in series such that sparge exhaust from a first fermentor or a set of fermentors in the series is used as a gas supply for a second fermentor or set of fermentors in the series.